文献类型：期刊文献

英文题名：Stepwise genetic modification for efficient expression of heterologous proteins in Aspergillus nidulans

作者：Yan, Qin[1];Han, Laichuang[1];Liu, Zhongmei[1,2];Zhou, Shengmin[3];Zhou, Zhemin[1,2]

机构：[1]Jiangnan Univ, Sch Biotechnol, Wuxi, Jiangsu, Peoples R China;[2]Jiangnan Univ, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China;[3]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China

年份：2023

卷号：107

期号：22

起止页码：6923

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20233814733384);WOS:【SCI-EXPANDED(收录号:WOS:001066997600002)】；

基金：This work is supported by the National Natural Science Foundation of China (32201034), the Natural Science Foundation of Jiangsu (BK20210470), China Postdoctoral Science Foundation (2021M701461), the International S&T Innovation Cooperation programme (2017YFE0129600), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (no. 111-2-06).

语种：英文

外文关键词：Aspergillus nidulans; Superior chassis cell; Mycelial morphology; Protein secretory pathway; Human interleukin-6

摘要：Filamentous fungi are widely used in food fermentation and therapeutic protein production due to their prominent protein secretion and post-translational modification system. Aspergillus nidulans is an important model strain of filamentous fungi, but not a fully developed cell factory for heterologous protein expression. One of the limitations is its relatively low capacity of protein secretion. To alleviate this limitation, in this study, the protein secretory pathway and mycelium morphology were stepwise modified. With eGFP as a reporter protein, protein secretion was significantly enhanced through reducing the degradation of heterologous proteins by endoplasmic reticulum-associated protein degradation (ERAD) and vacuoles in the secretory pathway. Elimination of mycelial aggregation resulted in a 1.5-fold and 1.3-fold increase in secretory expression of eGFP in typical constitutive and inducible expression systems, respectively. Combined with these modifications, high secretory expression of human interleukin-6 (HuIL-6) was achieved. Consequently, a higher yield of secretory HuIL-6 was realized by further disruption of extracellular proteases. Overall, a superior chassis cell of A. nidulans suitable for efficient secretory expression of heterologous proteins was successfully obtained, providing a promising platform for biosynthesis using filamentous fungi as hosts.